Rat beta-NGF DuoSet ELISA
Rat beta-NGF DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat beta-NGF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: beta-NGF
Nerve growth factor (NGF) was initially isolated in the mouse submandibular gland over three decades ago as a 7S complex composed of three non- covalently linked subunits, alpha, beta, and gamma. It is now known that both the alpha and gamma subunits of NGF are members of the kallikrein family of serine proteases, while the beta subunit, called beta-NGF or 2.5S NGF, exhibits all the biological activities ascribed to NGF. The human protein shares approximately 90% identity at the amino acid level with both mouse and rat beta-NGF.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Rat beta-NGF DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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Artificial nerve graft constructed by coculture of activated Schwann cells and human hair keratin for repair of peripheral nerve defects
Authors: HJ Qin, H Li, JZ Chen, KR Zhang, XQ Zhao, JQ Qin, B Yu, J Yang
Neural regeneration research, 2023-05-01;18(5):1118-1123.
Species: Rat
Sample Types: Cell Culture Supernates
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Stereological Evidence of Non-Selective Hippocampal Neurodegeneration, IGF-1 Depletion, and Behavioral Deficit following Short Term Bilateral Adrenalectomy in Wistar Rats
Authors: N Hamadi, ÖG Deniz, ASA Issa, AUS Islam, N Amir, ST Minhas, N Madjid, F Khelifi-To, S Kaplan, A Adem
Biomolecules, 2022-12-22;13(1):.
Species: Rat
Sample Types: Tissue Homogenates
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Peripheral Nerve Regeneration-Adipose-Tissue-Derived Stem Cells Differentiated by a Three-Step Protocol Promote Neurite Elongation via NGF Secretion
Authors: S Klein, A Siegmund, A Eigenberge, V Hartmann, F Langewost, N Hammer, A Anker, K Klein, C Morsczeck, L Prantl, O Felthaus
Cells, 2022-09-15;11(18):.
Species: Rat
Sample Types: Cell Culture Supernates
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Stimulation of Neurite Outgrowth Using Autologous NGF Bound at the Surface of a Fibrous Substrate
Authors: MR Casanova, RL Reis, A Martins, NM Neves
Biomolecules, 2021-12-24;12(1):.
Species: Rat
Sample Types: Plasma
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Dental pulp stem cells stimulate neuronal differentiation of PC12 cells
Authors: N Sultan, LE Amin, AR Zaher, ME Grawish, BA Scheven
Neural regeneration research, 2021-09-01;16(9):1821-1828.
Species: Rat
Sample Types: Cell Culture Supernates
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TGF-beta regulates nerve growth factor expression in a mouse intervertebral disc injury model
Authors: Y Yokozeki, K Uchida, A Kawakubo, M Nakawaki, T Okubo, M Miyagi, G Inoue, M Itakura, H Sekiguchi, M Takaso
Bmc Musculoskeletal Disorders, 2021-07-23;22(1):634.
Species: Mouse
Sample Types: Cell Culture Supernates
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Quercetin protects against diabetic retinopathy in rats by inducing heme oxygenase-1 expression
Authors: GR Chai, S Liu, HW Yang, XL Chen
Neural regeneration research, 2021-07-01;16(7):1344-1350.
Species: Rat
Sample Types: Cell Culture Supernates
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Microenvironment-responsive immunoregulatory electrospun fibers for promoting nerve function recovery
Authors: K Xi, Y Gu, J Tang, H Chen, Y Xu, L Wu, F Cai, L Deng, H Yang, Q Shi, W Cui, L Chen
Nat Commun, 2020-09-09;11(1):4504.
Species: Rat
Sample Types: Protein
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Insulin treatment protects the brain against neuroinflammation by reducing cerebral cytokines and modulating mitochondrial function
Authors: P Bortoluzzi, D Casagrande, N Dos Santos, K Ugioni Sim, C Mendes, ME Anastácio, PC Lock Silve, A Pastoris M
Brain Res. Bull., 2019-04-16;0(0):.
Species: Rat
Sample Types: Tissue Homogenates
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Nerve growth factor continuously elevates in a rat rotator cuff tear model
Authors: N Nagura, T Kenmoku, K Uchida, M Nakawaki, G Inoue, M Takaso
J Shoulder Elbow Surg, 2018-08-23;28(1):143-148.
Species: Rat
Sample Types: Tissue Homogenates
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Lifelong environmental enrichment in the absence of exercise protects the brain from age-related cognitive decline
Authors: AM Birch, ÁM Kelly
Neuropharmacology, 2018-04-07;0(0):.
Species: Rat
Sample Types: Tissue Culture Supernates
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Electroacupuncture and moxibustion promote regeneration of injured sciatic nerve through Schwann cell proliferation and nerve growth factor secretion
Authors: LN Hu, JX Tian, W Gao, J Zhu, FF Mou, XC Ye, YP Liu, PP Lu, SJ Shao, HD Guo
Neural Regen Res, 2018-03-01;13(3):477-483.
Species: Rat
Sample Types: Serum
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Isoflurane and the analgesic effect of acupuncture and electroacupuncture in an animal model of neuropathic pain
Authors: LN Spezia Ada, R Vercelino, C de Oliveir, VL Scarabelot, A de Souza, LF Medeiros, SG Cioato, W Caumo, ILS Torres
J Acupunct Meridian Stud, 2018-02-06;0(0):.
Species: Rat
Sample Types: Tissue Homogenates
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Therapeutic Benefit for Late, but Not Early, Passage Mesenchymal Stem Cells on Pain Behaviour in an Animal Model of Osteoarthritis
Authors: V Chapman, H Markides, DR Sagar, L Xu, JJ Burston, P Mapp, A Kay, RH Morris, O Kehoe, AJ El Haj
Stem Cells Int, 2017-12-24;2017(0):2905104.
Species: Rat
Sample Types: Serum
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Malathion increases apoptotic cell death by inducing lysosomal membrane permeabilization in N2a neuroblastoma cells: a model for neurodegeneration in Alzheimer's disease
Authors: R Venkatesan, YU Park, E Ji, EJ Yeo, SY Kim
Cell Death Discov, 2017-04-24;3(0):17007.
Species: Rat
Sample Types: Cell Culture Supernates
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A biphenyl derivative from the twigs of Chaenomeles speciosa
Authors: WS Suh, KJ Park, DH Kim, L Subedi, SY Kim, SU Choi, KR Lee
Bioorg. Chem., 2017-04-11;72(0):156-160.
Species: Rat
Sample Types: Cell Culture Supernates
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Time-Dependent Nerve Growth Factor Signaling Changes in the Rat Retina During Optic Nerve Crush-Induced Degeneration of Retinal Ganglion Cells
Authors: LA Mesentier-, S De Nicol•, P Rosso, LA De Vitis, V Castoldi, L Leocani, R Mendez-Ote, MF Santiago, P Tirassa, P Rama, A Lambiase
Int J Mol Sci, 2017-01-05;18(1):.
Species: Rat
Sample Types: Tissue Homogenates
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Incorporation of chitosan microspheres into collagen-chitosan scaffolds for the controlled release of nerve growth factor.
Authors: Zeng, Wen, Rong, Mengyao, Hu, Xueyu, Xiao, Wei, Qi, Fengyu, Huang, Jinghui, Luo, Zhuojing
PLoS ONE, 2014-07-01;9(7):e101300.
Species: Rat
Sample Types: Cell Culture Supernates
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Rab7 silencing prevents mu-opioid receptor lysosomal targeting and rescues opioid responsiveness to strengthen diabetic neuropathic pain therapy.
Authors: Mousa S, Shaqura M, Khalefa B, Zollner C, Schaad L, Schneider J, Shippenberg T, Richter J, Hellweg R, Shakibaei M, Schafer M
Diabetes, 2012-12-10;62(4):1308-19.
Species: Rat
Sample Types: Tissue Homogenates
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Electrical stimulation of sympathetic neurons induces autocrine/paracrine effects of NGF mediated by TrkA.
Authors: Saygili E, Schauerte P, Kuppers F, Heck L, Weis J, Weber C, Schwinger RH, Hoffmann R, Schroder JW, Marx N, Rana OR
J. Mol. Cell. Cardiol., 2010-02-02;49(1):79-87.
Species: Rat
Sample Types: Cell Culture Supernates
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The multi-herbal medicine Gongjin-dan enhances memory and learning tasks via NGF regulation.
Authors: Moon E, Her Y, Lee JB, Park JH, Lee EH, Kim SH, Oh MS, Jang CG, Kim SY
Neurosci. Lett., 2009-09-27;466(3):114-9.
Species: Rat
Sample Types: Cell Culture Supernates
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Anti-NGF-antibody administration as collyrium reduces the presence of NGF and enhances the expression of VEGF in the retina, lacrimal gland and hippocampus.
Authors: Colafrancesco V, Cirulli F, Rossi S, Berry A, Aloe L
Neurosci. Lett., 2009-08-03;463(3):203-6.
Species: Rat
Sample Types: Tissue Homogenates
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Low-frequency electro-acupuncture reduces the nociceptive response and the pain mediator enhancement induced by nerve growth factor.
Authors: Aloe L, Manni L
Neurosci. Lett., 2008-11-07;449(3):173-7.
Species: Rat
Sample Types: Serum
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Effects of nicergoline on corneal epithelial wound healing in rat eyes.
Authors: Kim SY, Choi JS, Joo CK
Invest. Ophthalmol. Vis. Sci., 2008-10-03;50(2):621-5.
Species: Rat
Sample Types: Cell Culture Supernates
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Eosinophil-nerve interactions and neuronal plasticity in rat gut associated lymphoid tissue (GALT) in response to enteric parasitism.
Authors: O'Brien LM, Fitzpatrick E, Baird AW, Campion DP
J. Neuroimmunol., 2008-05-20;197(1):1-9.
Species: Rat
Sample Types: Tissue Homogenates
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Maternal poly I:C exposure during pregnancy regulates TNF alpha, BDNF, and NGF expression in neonatal brain and the maternal-fetal unit of the rat.
Authors: Gilmore JH, Jarskog LF, Vadlamudi S
J. Neuroimmunol., 2004-11-24;159(1):106-12.
Species: Rat
Sample Types: Tissue Homogenates
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Intracerebral transplantation of carotid body in rats with transient middle cerebral artery occlusion.
Authors: Yu G, Xu L, Hadman M, Hess DC, Borlongan CV
Brain Res., 2004-07-23;1015(1):50-6.
Species: Rat
Sample Types: Tissue Homogenates
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Intravenous administration of marrow stromal cells (MSCs) increases the expression of growth factors in rat brain after traumatic brain injury.
Authors: Mahmood A, Lu D, Chopp M
J. Neurotrauma, 2004-01-01;21(1):33-9.
Species: Rat
Sample Types: Tissue Homogenates
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